臺灣博碩士論文加值系統

大蒜為世界各地常用之香辛調味料，且臺灣的大蒜品質優良，但近年來常受到進口產品競爭、環境氣候變遷或產量失衡等影響，造成大蒜供需失調、價格波動劇烈及影響蒜農產業的發展。開發具吸引力的加工產品如黑蒜保健產品，不失是解決上述問題的良方。雖然近年來有一些黑蒜加工產品上市，但其製程都需長時間，且鮮少有完整的研究數據探討黑蒜發酵製程與產品品質之相關性。故本研究以臺灣蒜頭最大產區-雲林縣莿桐鄉的大蒜為原料，在相對溼度70%下以溫度40℃和70℃分別進行發酵24天，並探討發酵期間蒜頭的理化性質及感官評估之變化，同時研究發酵過程其乙醇及水萃取物之抗氧化能力、抑菌能力及抗發炎活性之更迭。研究結果顯示，大蒜一般成分分析中以總醣所占比例最高，其含量為76.69%，其次為粗蛋白18.42%。理化性質的變化以70℃發酵之產品較為明顯，在水含量、水活性、pH值、L值及b值皆隨著發酵天數的增加而下降，他們在第24天時有最低之測定值；反之褐變強度則隨著發酵時間的增加而遞增。然而40℃發酵產品只有輕度的褐變，所以在理化性質的變化不若70℃發酵產品顯著，故40℃發酵黑蒜產品的亮度及黃色度分析值較大。另外在抗氧化能力方面，清除DPPH自由基的能力，70℃產品優於40℃發酵之黑蒜，其中乙醇萃取物優於水萃取物，發酵後之乙醇萃取物各實驗組自由基清除率約為71.67-77.72%；反之，亞鐵離子螯合能力則以40℃發酵產品高於70℃，在發酵第24天之乙醇萃取物有最高之測定值，其值為90.10%；還原力以70℃發酵之水萃取物在第24天有最高之測定值，其值為2.24；總酚及類黃酮含量，在70℃發酵期間隨著發酵天數的增加其含量隨之上升，以發酵第24天水萃取物有最高之測定值，其分析值分別為33.07及5.41 (mg/ml)。然而抑菌能力則以新鮮大蒜的具有較強之抑菌活性，其中以水萃取對於Bacillus cereus ATCC 14579有最大之抑菌圈，其值為3.30 mm；40℃發酵之萃取物隨著發酵天數增加，其抑菌能力遞減，在70℃發酵部分在第8天就無法測得抑菌活性。在抗發炎試驗中，新鮮大蒜及黑蒜之萃取物對小鼠巨噬細胞RAW264.7皆不會降低細胞之存活率；在抑制一氧化氮(Nitric oxide；NO)生成部分以40℃發酵黑蒜能有效抑制NO的產生，且抑制能力顯著大於70℃及新鮮大蒜，在發酵第8天之乙醇萃取有最好之抑制能力，抑制率為36.65%；在抑制一氧化氮合成酶(inducible NOS；iNOS)的表現方面，不論40℃及70℃發酵之黑蒜皆能有效抑制iNOS表現量，且乙醇萃取物的抑制效果大於水萃取物，而在40℃發酵第8天有最好之抑制iNOS表現量，其值降低40.67%；發酵後各實驗組皆能有效抑制腫瘤壞死因子-α(tumor necrosis factor α；TNF-α)的生成。總之，黑蒜發酵過程中其理化性質與抗氧化能力的變化是息息相關的，而根據各項抗氧化能力指標中，黑蒜70℃發酵之乙醇萃取物在第16-24天其測定值已達平穩；而抗發炎能力分析中，發酵後皆能有效抑制一氧化氮及一氧化氮合成酶的生成，故在相對溼度70%及溫度70℃條件下經16-24天發酵即可成不錯的黑蒜產品，其對抗氧化及抗發炎能力皆明顯增加，比未發酵大蒜更具有保健功效。

Garlic is commonly used as spicy seasoning in the world, Taiwan has high quality garlic, but in recent years, Taiwan suffers the impacts including the competition of imports, environment climate change or yield imbalance, resulting in the maladjustment between garlic supply and demand, price fluctuations and influences the development of garlic agriculture. The good way to solve these problems is exploiting attractive processing products such as black garlic health care products. Although some garlic processing products have been released on the market in recent years, but it takes a long time to ferment and it is rare in completed researches on studying in the correlation between the black garlic fermentation process and the product quality. Therefore, in this study, we use the garlic from Citong Township, Yun Lin County which is the largest production area of Taiwan as raw materials, carry on 24 days fermentation under the relative humidity of 70% and temperature of 40℃ and 70℃, respectively, investigating the changes of their physicochemical properties and sensory evaluation and also studying the variations of antioxidant ability, antibacterial activity and anti-inflammatory activity of their ethanol and water extracts during fermentation. The results demonstrated that total carbohydrate had the highest content ratio in garlic general components analysis which was 76.69%, followed by crude protein 18.42%. The fermentation products which temperature were 70℃ had largest changes in physicochemical properties, including the water content, water activity, pH value, L value and b value, were decreased with the increase in the number of fermentation days, they had the lowest measured value at 24 days. On the contrary, the browning intensity increased with the increase of fermentation time. However, the fermentation products of 40℃ had only slight browning, so the changes in physicochemical properties were not as obvious as the fermentation products of 70℃, thus, they had larger analysis value in brightness and yellowness. In the aspect of antioxidant capacity, 70℃ fermentation of black garlics products were statistically significant better than 40℃ in the ability to remove DPPH free radicals, and their ethanol extracts were better than water extracts which had 71.67-77.72% of scavenging percentage. On the other hand, ferrous ions chelating ability of 40℃ fermentation products were higher than 70℃, the ethanol extracts on the 24th day of fermentation had the highest measured value of 90.10%. The water extracts at 70℃ on 24th day had the highest reducing power which had the value of 2.24. The content of total polyphenols and flavonoids in 70℃ fermentation products increased with the rise in the number of fermentation days, especially in 24th day, had the highest value of 33.07 and 5.41 (mg / ml), respectively. Nevertheless, fresh garlic had the strongest antibacterial activity; particularly their water extract had the largest inhibition zone for Bacillus cereus ATCC 14579 with the value of 3.30 mm. Moreover, the antibacterial activity of the extract at 40℃ decreased progressively with the increase in the number of fermentation days. In 70℃ fermentation, the antibacterial activity could not be detected on the 8th day. The anti-inflammatory test illustrated that extracts of fresh or black garlics were not declined the survival rate of mouse macrophage RAW264.7 cells. Furthermore, the black garlics in fermentation temperature of 40℃ could effectively inhibit the productions of nitric oxide (NO), and the inhibition ability were significantly higher than 70℃ and fresh garlic, the ethanol extracts on 8th day had the best inhibition ability with the inhibition rateis 63.35%. We further investigated the expression of nitric oxide synthase (iNOS), the results showed that the black garlics in fermentation temperature of 40℃ and 70℃, both had the capacity on inhibiting the expression of iNOS, and the ethanol extracts were more effective than the water extracts. However, the fermentation temperature of 40℃ on 8th day had the best capacity on inhibiting the expression of iNOS with decreasing the value to 40.67%.Besides that, TNF-α production was inhibited by all experimental groups after fermentation. In conclusion, the antioxidant ability of black garlic is closely related to their physicochemical properties during fermentation, according to the indexes of antioxidant ability, the ethanol extracts of black garlics fermenting at 70 ℃ have stable level on the 16th to 24th day; while in the anti-inflammatory ability analysis, the fermented black garlics could effectively inhibit the productions of nitric oxide and nitric oxide synthase. Hence, carrying on fermentation for 16-24 days under relative humidity of 70% and temperature of 70℃, a batch of good quality black garlic can be obtained, their antioxidant and anti-inflammatory properties are better than fresh garlic and they possess health benefits.

目錄Ⅲ
表目錄Ⅴ
圖目錄Ⅵ
摘要Ⅶ
AbstractⅨ
縮寫表XI
第一章 前言1
第二章 文獻探討3
第一節大蒜3
一、大蒜簡介3
二、大蒜的機能性成分6
三、大蒜的保健功效8
四、大蒜的加工應用10
第二節黑蒜12
一、黑蒜簡介12
二、黑蒜加工條件13
三、黑蒜的化學成分14
第三章 材料與方法16
第一節實驗架構16
第二節材料17
第三節化學試劑17
第四節儀器設備19
第五節實驗方法20
一、一般成分分析21
二、色澤分析21
三、水活性測定21
四、pH值測定21
五、褐變強度分析22
六、萃取方法22
七、清除DPPH 自由基能力之測定22
八、亞鐵離子螯合能力之測定22
九、還原力之測定23
十、總酚含量分析23
十一、類黃酮含量分析23
十二、微生物之培養23
十三、抑菌圈及最小抑菌濃度24
十四、細胞之培養24
十五、細胞存活率試驗24
十六、降發炎物質NO試驗25
十七、RNA萃取25
十八、反轉錄製備成cDNA26
十九、即時定量聚合酵素連鎖反應(Q-PCR)26
二十、感官評估26
二十一、統計分析27
第四章 結果與討論28
第一節大蒜的一般成分分析28
第二節黑蒜發酵24天期間理化性質之分析28
第三節黑蒜發酵24天期間感官評估之分析31
第四節 黑蒜發酵24天期間抗氧化能力之分析31
第五節黑蒜發酵24天期間抗氧化能力相關性之分析34
第六節黑蒜發酵24天期間抑菌能力之分析34
第七節黑蒜發酵24天期間抗發炎能力之分析36
第五章 結論38
參考文獻40
附表48
附圖54




表目錄
表一、大蒜的一般成分分析48
表二、黑蒜發酵24天期間色澤之變化49
表三、黑蒜發酵24天期間感官評估之變化50
表四、黑蒜發酵24天期間相關性之分析51
表五、黑蒜發酵24天期間抑菌性之變化52
表六、黑蒜發酵24天期間最小抑菌濃度之變化53



圖目錄
圖一、台灣地區常見大蒜品種之照片54
圖二、黑蒜發酵24天期間之照片56
圖三、黑蒜發酵24天期間水含量之變化58
圖四、黑蒜發酵24天期間水活性之變化60
圖五、黑蒜發酵24天期間pH值之變化62
圖六、黑蒜發酵24天期間褐變強度之變化64
圖七、黑蒜發酵24天期間清除DPPH自由基能力之變化66
圖八、黑蒜發酵24天期間亞鐵離子螯合能力之變化68
圖九、黑蒜發酵24天期間還原力之變化70
圖十、黑蒜發酵24天期間總酚之變化72
圖十一、黑蒜發酵24天期間類黃酮之變化74
圖十二、黑蒜發酵24天期間巨噬細胞RAW264.7細胞存活率之變化76
圖十三、黑蒜發酵24期間抑制巨噬細胞RAW264.7一氧化氮(NO)表現之變化78
圖十四、黑蒜發酵24天期間抑制巨噬細胞RAW264.7 iNOS生成之變化80
圖十五、黑蒜發酵24天期間抑制巨噬細胞RAW264.7 TNF-α生成之變化82

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